Femoral Bone Microstructure Research Articles

Long-Term Cola Intake Does Not Cause Evident Pathological Alterations in the Femoral Bone Microstructure: An Animal Study in Adult Mice.

Short-term animal experiments and association studies in humans have shown that cola intake may have a detrimental impact on bone mineral density (BMD); however, other bone parameters have not been investigated. This study examined the effects of long-term cola consumption on the femoral bone microstructure using adult mice (n = 32) as an animal model, which were divided into water and cola groups depending on whether they received water or cola along with a standard rodent diet for 6 months. Micro-computed tomography revealed that cola intake did not significantly affect all measured parameters characterizing trabecular bone mass and microarchitecture, as well as cortical microarchitecture and geometry in both sexes, although a slight deterioration of these parameters was noted. Cola consumption also resulted in a slightly, statistically insignificant worsening of bone mechanical properties. In contrast to female mice, males receiving cola had a lower area of primary osteons' vascular canals. Nevertheless, long-term cola intake did not cause evident pathological alterations in the femur of adult mice, possibly due to a balanced diet and no restriction of physical activity. Therefore, the adverse effects of cola consumption on BMD, the only bone parameter studied so far, may be caused by other risk and lifestyle factors.

The effects of eggshell calcium (Biomin H® ) and its combinations with alfacalcidol (1α-hydroxyvitamin D3) and menaquinone-7 (vitamin K2) on ovariectomy-induced bone loss in a rat model of osteoporosis.

The purpose of this study was to investigate the impact of eggshell calcium (Biomin H® dietary supplement) and its combinations with alfacalcidol (1α-hydroxyvitamin D3 ) and menaquinone-7 (vitamin K2 ) on ovariectomy-induced bone loss in rats. Adult female rats (n=48) were divided into 6 groups of 8 individuals each: sham-operated rats (SHAM); ovariectomized (OVX) rats untreated; OVX rats treated with Biomin H® (BIO); OVX rats simultaneously receiving Biomin H® , vitamin D3 (BIO+D3 ); OVX rats simultaneously treated with Biomin H® , vitamin K2 (BIO+K2 ) and OVX rats treated with Biomin H® , vitamin D3 , vitamin K2 (BIO+D3 +K2 ) during 8weeks. Biochemical parameters, bone mineral density (BMD), bone mineral content (BMC) and femoral bone microstructure were determined. Plasma calcium and phosphate were increased in BIO+D3 and BIO+D3 +K2 groups as compared to OVX. Alkaline phosphatase was elevated in OVX, BIO versus SHAM, BIO+D3 +K2 groups. When compared to OVX group, decreased urine deoxypyridinoline was observed in all treated groups and femoral BMD, BMC were higher in BIO, BIO+D3 , BIO+D3 +K2 groups. The BIO+K2 rats had similar densitometrical values than OVX individuals. Microcomputed tomography revealed increased trabecular relative bone volume (due to an increase in trabecular number) in BIO+D3 , BIO+D3 +K2 as compared to OVX. The higher relative bone volume in BIO+D3 , BIO+D3 +K2 groups was also accompanied by an increase in bone surface. In the cortical bone, an enhanced periosteal bone apposition was identified in BIO, BIO+D3 , BIO+K2 , BIO+D3 +K2 groups. The rats from BIO+D3 +K2 group had a higher area of primary osteon's vascular canals. In BIO+D3 , BIO+K2 , BIO+D3 +K2 groups, an increased area of secondary osteons was determined in comparison with OVX. Our results indicate the beneficial effect of triple application of Biomin H® , vitamin D3 , vitamin K2 , as well as simultaneous administration of Biomin H® , vitamin D3 on the inhibition of ovariectomy-induced bone loss in a rat model of osteoporosis.

Effect of the mixture of mulberry leaf powder and KGM flour on promoting calcium absorption and bone mineral density in vivo.

In this paper, mulberry leaf powder (MLP) and konjac glucomannan (KGM) flour were used as raw materials, and animal experiments were designed to evaluate the effects of a mixture of MLP and KGM on bone density. The femoral bone microstructure of mice and pathological changes were observed by using micro-computed tomography) and haematoxylin and eosin (HE) staining methods, respectively. A three-point bending test was used to determine the biomechanical properties of the femur. Results indicated that the calcium content of MLP was high, reaching 16 148.5 mg kg-1 , and the total proportion of water-soluble calcium, calcium pectinate, and calcium carbonate accounted for about 60% of the total calcium content. Serum alkaline phosphatase (AKP) activity was significantly lower, and serum calcium content was significantly higher (P < 0.05), in the MLP + KGM group (KM) than in the low-calcium control group, whereas no significant difference (P > 0.05) was found for serum phosphorus content. KM had a longer femur length, a higher bone mineral density (BMD) (P > 0.05), and significantly greater femur diameter, dry weight, index and bone calcium content (P < 0.05). However, these parameters were not significantly different from those of the calcium carbonate control group (P > 0.05). The results indicate that the MLP/KGM mixture can reduce the high rate of bone turnover and the corresponding loss of bone mass caused by calcium deficiency and is thus effective in enhancing bone density. © 2019 Society of Chemical Industry.

Single and simultaneous effects of acrylamide and ethanol on bone microstructure of mice after one remodeling cycle

BackgroundThis study aimed to examine femoral bone microstructure of mice after single and simultaneous administration to acrylamide and ethanol since both substances are often consumed separately and/or together by humans. Interactive effects of these toxins were analysed after one remodeling cycle.MethodsTwenty clinically healthy adult mice were randomly divided into four groups following 2 weeks administration of toxins: A group - mice were fed with acrylamide (40 mg/kg bw); E group - mice were ethanol-fed (15% ethanol); AE group - mice were simultaneously fed with both toxins, and a C group – control (without acrylamide and/or ethanol supplementation). Generally, 2D and 3D imaging methods were used to determine cortical and trabecular bone tissues microstructure. Biochemical analyses of plasma parameters were also realized using commercially available ELISA tests and spectrophotometrically.ResultsSingle and simultaneous exposure to acrylamide and ethanol affected only cortical bone microstructure. No significant changes in trabecular bone morphometry were detected among all groups. In mice from the A group, increased endocortical remodeling associated with a higher level of serum calcium and vasoconstriction of primary osteon’s vascular canals (POVC) were identified. On the contrary, increased cortical porosity consistent with a decreased relative bone volume, bone mineral density (BMD) and lower levels of alkaline phosphatase (ALP), glutathione (GSH), calcium in plasma and also with vasodilation of POVC were observed in the E group. In the AE group, the highest density of secondary osteons associated with a lower BMD and decreased levels of ALP, GSH were documented. The parameters of POVC and Haversian canals approximated to the C group. In addition, single and simultaneous exposure to both toxins caused liver disease consistent with a higher values of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in plasma of all experimental groups.ConclusionsSingle administration to acrylamide and ethanol had negative effects on cortical bone structure of mice after one remodeling cycle. However, we identified possible antagonistic impact of these toxins on the structure of the cortical bone.

Palaeohistology and life history evolution in cave bears, Ursus spelaeus sensu lato.

The abundance of skeletal remains of cave bears in Pleistocene deposits can offer crucial information on the biology and life history of this megafaunal element. The histological study of 62 femora from 23 different European localities and comparisons with specimens of five extant ursid species revealed novel data on tissue types and growth patterns. Cave bear’s femoral bone microstructure is characterized by a fibrolamellar complex with increasing amounts of parallel-fibered and lamellar bone towards the outer cortex. Remodelling of the primary bone tissue initially occurs close to the perimedullary margin of the bone cortex around the linea aspera. Although similar histological traits can be observed in many extant bear species, the composition of the fibrolamellar complex can vary greatly. Cave bears reached skeletal maturity between the ages of 10 and 14, which is late compared to other bear species. There is a significant correlation between altitude and growth, which reflects the different body sizes of cave bears from different altitudes.

A POSITIVE DOSE-RESPONSE OF QUERCETIN ON COMPACT BONE MICROSTRUCTURE OF MALE RABBITS

Quercetin is one of the most widely distributed flavonoids in plants which should have a broad range of significant health promoting properties. Its effect on bone microstructure of males has not been sufficiently investigated. Therefore, our study was aimed to determine the impact of quercetin on femoral bone microstructure in 5-month-old male rabbits. Nine rabbits were intramuscularly injected with following doses of quercetin: 10 μg/kg body weight (bw) (E1 group; n=3), 100 μg/kg bw (E2 group; n=3) and 1000 μg/kg bw (E3 group; n=3) for 90 days, 3 times per week. Three rabbits without quercetin administration served as a control group. An accelerated bone resorption at the endosteal surface and intensive periosteal bone apposition were observed in all rabbits administered quercetin. However, the most evident changes such as a thicker layer of primary vascular longitudinal bone tissue under periosteum and a lower density of secondary osteons were identified in males from the E3 group. Morphometrical evaluations of the compact bone showed significantly decreased sizes of primary osteons’ vascular canals and secondary osteons in rabbits exposed to the highest dose of quercetin. Sizes of secondary osteons were also lower in males from the E2 group. Our results demonstrate that quercetin has a positive dose-response on compact bone microstructure in male rabbits.

Acrylamide-induced changes in femoral bone microstructure of mice.

Acrylamide (AA) is one of the most common toxins in foods. Its effect on bone microstructure has not been investigated. The aim of our study was to analyze the impact of acute exposure to AA on femoral bone microstructure in mice. Adult animals were treated perorally with 2 doses of AA (E1 group, 1 mg/kg b.w.) in a 24-h period and with 3 doses of AA (E2 group, 1 mg/kg b.w.) in a 48-h period. Mice exposed to AA had smaller sizes of primary osteon's vascular canals. Secondary osteons were significantly smaller in mice from E2 group; however their increased number (from 38 % to 77 %) was identified in both E1 and E2 groups. In these groups, a higher number of resorption lacunae (from 100 % to 122 %) was also found. The values for bone volume, trabecular number were increased and that for trabecular separation was decreased in mice administered AA. Significantly higher value of bone surface was observed in mice from E1 group whereas trabecular thickness was increased in E2 group. The effect of AA on microstructure of compact and trabecular bone tissues is different. In our study, one dose of AA was used and acute effects of AA were investigated. Therefore, further studies are needed to study mechanisms by which AA acts on bone.

The Effect of Subacute Exposure to Acrylamide on Femoral Bone Microstructure in Laboratory Mice

The Effect of Subacute Exposure to Acrylamide on Femoral Bone Microstructure in Laboratory Mice

Histological analysis of femoral bones in rabbits administered by amygdalin

Cyanogenic glycosides are present in several economically important plant foods. Amygdalin, one of the most common cyanoglucoside, can be found abundantly in the seeds of apples, bitter almonds, apricots, peaches, various beans, cereals, cassava and sorghum. Amygdalin has been used for the treatment of cancer, it shows killing effects on cancer cells by release of cyanide. However, its effect on bone structure has not been investigated to date. Therefore, the objective of this study was to determine a possible effect of amygdalin application on femoral bone microstructure in adult rabbits. Four month old rabbits were randomly divided into two groups of three animals each. Rabbits from E group received amygdalin intramuscularly at a dose 0.6 mg.kg-1 body weight (bw) (group E, n = 3) one time per day during 28 days. The second group of rabbits without amygdalin supplementation served as a control (group C, n = 3). After 28 days, histological structure of femoral bones in both groups of rabbits was analysed and compared. Rabbits from E group displayed different microstructure in middle part of the compact bone and near endosteal bone surface. For endosteal border, an absence of the primary vascular longitudinal bone tissue was typical. This part of the bone was formed by irregular Haversian and/or by dense Haversian bone tissues. In the middle part of substantia compacta, primary vascular longitudinal bone tissue was observed. Cortical bone thickness did not change between rabbits from E and C groups. However, rabbits from E group had a significantly lower values of primary osteons' vascular canals and secondary osteons as compared to the C group. On the other hand, all measured parameters of Haversian canals did not differ between rabbits from both groups. Our results demonstrate that intramuscular application of amygdalin at the dose used in our study affects femoral bone microstructure in rabbits.

Quercetin-induced changes in femoral bone microstructure of adult male rabbits

Flavonoids are a group of plant metabolites with antioxidant effects. One of the most abundant flavonoids in the human diet is quercetin. It is found widely in fruits, vegetables and has a lot of beneficial effects on human health. Quercetin has a positive pharmacological effect on bone metabolism and it prevents the organism against bone loss. However, its impact on the size of basic structural units of the compact bone is still unknown. Therefore, the aim of present study was to investigate the impact of the quercetin on femoral bone microstructure in 5-month-old male rabbits. Five rabbits of Californian broiler line were randomly divided into two groups. In the experimental group (E group; n=3), animals were intramuscularly injected with quercetin at dose 1000 μg.kg-1 body weight (bw) for 90 days, 3 times per week. Two rabbits without quercetin administration served as a control group (C group). According to our results, intramuscular application of quercetin had an insignificant effect on cortical bone thickness in male rabbits. In these rabbits, changes in qualitative histological characteristics were present in the middle part of the compacta, where primary vascular longitudinal bone tissue was present and expanded there from the periosteum. Also, a lower number of secondary osteons was found in these animals. From the histomorphometrical point of view, significantly decreased sizes of primary osteons' vascular canals and secondary osteons (p &lt;0.05) were found in rabbits administered by quercetin. Our findings indicate that subchronic administration of quercetin at the dose used in our study had considerable impact on both qualitative and quantitative histological characteristics of the compact bone in adult male rabbits.

Structural changes in femoral bone microstructure of female rabbits after intramuscular administration of quercetin.

BackgroundQuercetin is one of the best known flavonoids being present in a variety of fruits and vegetables. It has cardioprotective, anticarcinogenic, antioxidant, anti-inflammatory and antiapoptotic properties. Some studies suggest that quercetin has protective effects on bone. However, its influence on qualitative and quantitative histological characteristics of compact bone is still unknown. In our study, 12 clinically healthy five-month-old female rabbits were divided into four groups of three animals each. Quercetin was applied intramuscularly in various concentrations; 10 µg/kg body weight (bw) in the E1 group, 100 µg/kg bw in the E2 group, and 1000 µg/kg bw in the E3 group for 90 days, 3 times per week. Three rabbits without exposure to quercetin served as a control (C) group. Differences in femoral bone microstructure among groups were evaluated.ResultsQualitative histological characteristics of compact bone differed between rabbits from the E1 and E2 groups. Primary vascular longitudinal bone tissue was not found in some areas near the endosteal surface due to increased endocortical bone resorption. In addition, periosteal border of rabbits from the E1 group was composed of a thicker layer of primary vascular longitudinal bone tissue than in the other groups. In all groups of rabbits administered quercetin, a lower density of secondary osteons was observed. Histomorphometrical evaluations showed significantly decreased sizes of the primary osteons’ vascular canals in individuals from the E1 and E2 groups. Secondary osteons were significantly smaller in rabbits from the E1, E2, E3 groups when compared to the C group. Cortical bone thickness was significantly increased in females from the E1 and E2 groups.ConclusionsThe results indicate that quercetin has not only a positive dose–response on qualitative and quantitative histological characteristics of the compact bone of female rabbits as it would be expected.

Sex-related variations in bone microstructure of rabbits intramuscularly exposed to patulin.

BackgroundPatulin, a toxic mold metabolite, has been found as natural contaminant of processed fruits, most notably apples, apple juices and other apple-based products. A number of adverse health effects in humans and animals are associated with patulin intoxication. The current study was performed to analyse possible toxic effects of patulin on femoral bone microstructure in adult rabbits in detail. Fourteen clinically healthy four-month-old rabbits of both sexes (6 males and 8 females) were included in the study. Animals from the experimental groups (group E♂, n = 3; group E♀, n = 4) were injected intramuscularly with patulin at dose 10 μg/kg body weight two times a week for 28 days. The dose of patulin was estimated based on the maximum permitted level of patulin for apple products intended for infants and young children. Three males and four females without patulin administration served as controls (groups C♂ and C♀). Cortical bone thickness and qualitative and quantitative histological characteristics of compact bone tissue were investigated.ResultsIntramuscular applications of patulin significantly increased the thickness of cortical bone in both sexes of rabbits. In patulin-exposed males, an absence of primary vascular longitudinal bone tissue near the endosteal border was observed, which could be associated with intensive bone remodeling. Femoral diaphyses of females displayed a lower number of secondary osteons in the middle part of the substantia compacta, and occurrence of the osteons near the periosteum. This could indicate alterations in bone turnover. Histomorphometrical evaluations showed significantly increased sizes of the primary osteons’ vascular canals (P < 0.05) in males exposed to patulin possibly due to mycotoxin-induced increased levels of testosterone.ConclusionsThis study demonstrates significant impact of intramuscular application of patulin on bone microstructure in adult rabbits. Moreover, we have found that the effects of patulin on qualitative and quantitative histological characteristics of compact bone are sex-dependent.

Osteohistological Evidence for Determinate Growth in the American Alligator

Abstract An external fundamental system (EFS) is a form of bone microstructure present in the outermost cortex of long bones in animals that have attained skeletal maturity. It indicates an effective cessation of any significant periosteal growth (i.e., growth in circumference or girth). Although an EFS has been noted in several reptile taxa, the idea that reptiles grow continually throughout their lives remains popular. Examination of femoral bone microstructure from captive American Alligators (Alligator mississippiensis) reveals parallel-fibered tissue terminating periosteally in an EFS, thus confirming determinate growth in another reptile taxon. The results of this study have several important implications for both modern and fossil tetrapods: first, because many birds, nonavian dinosaurs, pterosaurs, and basal pseudosuchians all produce an EFS, it can be concluded that determinate growth is a shared characteristic of Archosauria; second, because the captive alligators were not senescent, an EFS shou...

Effect of bioactive compounds in lactobacilli‐fermented soy skim milk on femoral bone microstructure of aging mice

Soy skim milks fermented with lactobacilli contain various phytochemicals such as isoflavones and peptides. We used lactobacilli-fermented soy skim milk as a nutritional supplement for 6 weeks to investigate its anti-osteoporosis effect in 13-month-old female BALB/c aging mice. Freeze-dried powder of soy skim milk fermented by Lactobacillus paracasei subsp. paracasei NTU 101 (NTU 101F) and L. plantarum NTU 102 (NTU 102F) were used in this study. The trabecular bone volumes in mice fed NTU 101F and NTU 102F increased by a factor of 3.48 and 2.16 compared with control values, respectively. The network density and thickness of distal metaphyseal trabecular bone in mice fed with NTU 101F and NTU 102F milks were significantly denser than that of control mice; moreover, the NTU 101F group had the largest resting area ratio and smallest resorbing area compared with other groups. The beneficial effect may due to isoflavones as well as higher amounts of polysaccharide and peptide in NTU 101F milk. The results suggest that dietary supplement with fermented soy skim milk can attenuate aging-induced bone loss in BALB/c mice and possibly lower the risk of osteopenia or osteoporosis in aging.